Preparation of chromatographic matrices by free radical addition ligand attachment to allyl groups

Abstract

Chromatographic matrices were produced by free radical addition reactions between immobilised allyl groups and suitable ligands. Efficient addition and high ligand densities were readily obtained with mercaptoacids, glutathione, mercaptoethanol and sodium bisulphite, using aqueous solvation. Addition of nitrogen containing thiol ligands other than glutathione was also demonstrated, although heat or radiation catalysis was required. Addition of cysteamine or mercaptoacids resulted in spacer arm derivatives, suitable for further ligand attachment chemistry. Mercaptoacetic acid and bisulphite attachment was used to prepare cation-exchange matrices. Glutathione derivatives were compared with matrices prepared conventionally, for affinity chromatography of glutathione transferase.